Fig. 1.

Manipulation of human Sirt1 expression levels in Caco-2 cells. a Confirmation by RT-PCR of Sirt1 overexpression [transient transfection with plasmid pCMV6-ENTRY-SIRT1 (Origene)] and knockdown (siRNA) by RT-qPCR. All data are for n = 18. b, c Confirmation by western blotting of Sirt1 overexpression (b) and knockdown (c). The specific Sirt1 and α-tubulin immunoreactive bands, with approximate molecular weights, are indicated. Approximately 10 μg of protein was loaded in each lane, and for each condition, three biological replicates are presented. In b, +SIRT1 indicates cells transfected with the Sirt1 overexpression construct. In c, siRNA (SIRT1) indicates cells transfected with one of two siRNAs (#1 or #2) targeted to Sirt1. All data are representative of multiple independent repeats of the procedures